Method and Apparatus for Producing a Stabilized Antimicrobial Non-toxic Electrolyzed Saline Solution Exhibiting Potential as a Therapeutic

ABSTRACT

An improved method and apparatus for producing a stable, non-toxic, antimicrobial electrolyzed saline solution with a broad range of anti-infective and therapeutic applications. The resulting electrolyzed saline solution exhibits a marked lack of toxicity upon intravenous, aspired, oral or topical application in mammals for therapeutic applications providing a broad platform, including topical disinfection, antimicrobial application, wound treatment, oxidative stress reduction and enhancement of immune function to better detect malfunctioning cells.

RELATED APPLICATIONS

This application is a divisional application of U.S. patent application Ser. No. 12/383,212, filed Mar. 20, 2009, which is a Continuation-in-Part of U.S. patent application Ser. No. 12/290,398, filed Oct. 30, 2008, which claims priority to U.S. Provisional Patent Application Ser. No. 61/001,010, filed Oct. 30, 2007, the entire contents all of which are herein incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field

This invention pertains to an electrolytic method and apparatus for producing electrolyzed saline redox-balanced solutions. More particularly, it pertains to a method and apparatus used to produce a stable, non-toxic, antimicrobial electrolyzed saline redox-balanced solution from pure saline or hypertonic saline (NaCl and H₂O), both referred to hereafter as saline solution, exhibiting anti-infective and immune-enhancing potential as a therapeutic employing a balanced mixture of chemically reduced and oxidized species including Hypochlorous acid (HOCl), Hypochlorites (OCl⁻, NaClO), dissolved Oxygen (O₂), Chlorine (Cl₂) and Hydrogen (H₂) gases, Hydrogen Peroxide (H₂O₂), Hydrogen ions (H⁺), Hypochloride (ClO) and corresponding amounts of Superoxides (*O₂ ⁻, HO₂), Ozone (O₃), Activated Hydrogen ions (H⁻), Chloride ions (Cl⁻), Hydroxides (NaOH, OH⁻), Singlet Oxygen (*O₂) and other forms of Reactive Oxygen Species (ROS) (*OCl, *HO⁻).

2. State of the Art

Electrolysis of saline solutions has long been used to produce antimicrobial solutions that are compatible with mammalian biology. Some examples include methods to produce chlorinated water, bleach and hydrogen peroxide. Typically, the methods and apparatus used to electrolyze these solutions employ ion-selective barriers between the electrodes in order to efficiently isolate the target molecules and eliminate unwanted byproducts. A fundamentally different method and apparatus for producing a non-toxic antimicrobial electrolyzed saline solution is disclosed in eight United States patents, and two Japanese patents and a Mexican patent based on these U.S. patents, all held by the applicant, covering various other applications for intravenous injected electrolyzed saline solution, the machinery that manufactures it, and the method by which it is manufactured. These U.S. Patents are as follows:

-   -   U.S. Pat. No. 5,334,383, Morrow, dated Aug. 2, 1994 entitled         “Electrically Hydrolyzed Salines as In Vivo Microbicides for         Treatment of Cardiomyopathy and Multiple Sclerosis.” This patent         covers a method of treating antigen related infections related         to cardiomyopathy and multiple sclerosis in humans and other         warm blooded animals. It does not cover the substance itself,         but covers a particular use of the substance. This method of         treatment includes the use of an electrolyzed saline solution in         conjunction with one or more modulating agents such as ascorbic         acid (Vitamin C), with or without concurrent colchicine, to         mimic or enhance the body's naturally occurring immune response         to bacterial, viral or fungal infection.     -   U.S. Pat. No. 5,507,932, dated Apr. 16, 1996 entitled “Apparatus         for Electrolyzing Fluids.” This patent covers equipment that         exposes a liquid solution to an electrical current, creating an         electrolyzed solution. This equipment may be used to produce an         electrolyzed saline solution, capable of killing bacterial,         viral and fungal agents, for use in medical applications such as         the treatment of antigen related infections in humans and other         warm blooded animals. This patent covers the equipment used to         produce the electrolyzed saline solution, not the substance         itself.     -   U.S. Pat. No. 5,560,816, Robinson, dated Oct. 1, 1996 entitled         “Method for Electrolyzing Fluids.” This patent covers a method         for electrolyzing fluids, by using specialized equipment to         expose liquid solutions to an electrical current. Saline, for         example, may be treated by this process to yield an electrolyzed         saline solution, capable of killing bacterial, viral and fungal         agents, for the treatment of antigen related infection in humans         and other warm blooded animals. This patent covers the method by         which the electrolyzed saline solution is produced, not the         substance itself.     -   U.S. Pat. No. 5,622,848, Morrow, dated Apr. 22, 1997 entitled         “Electrically Hydrolyzed Saline Solutions As Microbicides For In         Vitro Treatment Of Contaminated Fluids Containing Blood.” This         patent covers a method of treating whole blood and other blood         products with an electrolyzed saline solution to reduce         infection with bacterial, viral and fungal agents. This patent         covers a particular use of the electrolyzed saline solution, not         substance itself.     -   U.S. Pat. No. 5,674,537, Morrow, dated Oct. 7, 1997 entitled “An         Electrolyzed Saline Solution Containing Concentrated Amounts Of         Ozone And Chlorine Species.” This patent covers a specific         electrolyzed saline solution containing a regulated amount of         microbicidal agents including ozone and active chlorine species.         This solution is intended for use in the treatment of infections         in the body of humans and other warm blooded animals, or in         blood or blood products. This patent covers the electrolyzed         saline solution.     -   U.S. Pat. No. 5,731,008, Morrow, dated Mar. 24, 1998 entitled         “Electrically Hydrolyzed Salines as Microbicides.” This patent         covers a method of using a specific electrolyzed saline solution         containing a regulated amount of microbicidal agents including         ozone and active chlorine species for the treatment of microbial         infections, including HIV infection. The method includes         intravenous administration of the solution along with one or         more modulating agents such ascorbic acid (Vitamin C), with or         without concurrent colchicine. This patent covers a method for         using the electrolyzed saline solution, not the substance         itself.     -   U.S. Pat. No. 6,007,686, Welch et al, dated Dec. 28, 1999         entitled “System for Electrolyzing Fluids for Use as         Antimicrobial Agents.” This patent covers a system for         electrolyzing fluids, such as a saline solution, for use in         sterilizing dental and medical instruments and other health care         equipment. The patent covers the necessary equipment for         generating and circulating the electrolyzed saline solution         around the instruments to be sterilized, and includes specific         claims for equipment designed for use with dental drill hand         pieces and flexible tubing. This patent covers a process by         which the electrolyzed saline solution may be made for a         particular use, not the substance itself.     -   U.S. Pat. No. 6,117,285, Welch et al, dated Sep. 12, 2000         entitled “System for Carrying Out Sterilization of Equipment.”         This patent covers a system for cleaning and sterilizing medical         and dental instruments to prevent the spread of infection from         one patient to another. The covered system bathes the instrument         in an electrolyzed saline solution and causes the solution to         flow into and sterilize any openings in the equipment. It         includes specific claims for systems designed specifically for         the sterilization of dental drills and flexible tubing. This         patent covers a particular use of the electrolyzed saline         solution, not the substance itself.

Two Japanese and one Mexican patents provide corresponding coverage in those countries for several of the above U.S. patents. Applicants also have other pending applications with the US Patent and Trademark Office for patents on various treatment uses of the electrolyzed saline solution.

The above embodiments of these prior patents typically have produced measurably different variations of electrolyzed saline solution. Each variation, however, exhibited some antimicrobial action and many of these devices produced solutions with measurable amounts of the components (chlorine, pH, ozone, etc.) within the range of the disclosed regulated amounts. The resulting electrolyzed saline compositions, however, have not historically been satisfactorily consistent or controllable, specifically regarding the concentrations of Reactive Oxygen Species (ROS). In addition, these prior inventions could produce toxic chemicals (chlorates) in the process of electrolyzing the saline solution. Consequently, there is a need for an improved manufacturing method and apparatus, such as that described below, to consistently produce solutions suitable for therapeutic applications in humans and warm-blooded animals.

SUMMARY OF THE INVENTION

The improved method and apparatus described below provides an improved electrolyzing fluid containing regulated amounts of stable reactive oxygen species (ROS) particularly suited for stable, non-toxic antimicrobial applications and to aid the immune system in identifying and destroying malfunctioning cells. The invention comprises a method for making an electrolyzed saline solution for use as an in vivo treatment of a human or warm-blooded animal. Specifically, it comprises:

a. placing a saline solution having a saline concentration of at least about 0.15% within a container,

b. activating a fluid circulation device to maintain a flow of the saline solution between the electrode surfaces,

c. adjusting the temperature of the circulating saline at a preferred level to prevent production of chlorates and regulate the relative concentrations of resulting components,

d. placing in the saline solution an anode and a cathode associated with a power source, and

e. applying an effective voltage potential less than about thirty volts between the cathode and the anode sufficient to produce a balanced mixture of chemical redox balanced species including Hypochlorous acid (HOCl), Hypochlorites (OCl⁻, NaClO), dissolved Oxygen (O₂), Chlorine (Cl₂) and Hydrogen (H₂) gases, Hydrogen Peroxide (H₂O₂), Hydrogen ions (H⁺), Hypochloride (ClO) and corresponding amounts of Superoxides (*O₂ ⁻, HO₂), Ozone (O₃), Activated Hydrogen ions (H⁻), Chloride ions (Cl⁻), Hydroxides (NaOH, OH⁻), Singlet Oxygen (*O₂) and other forms of Reactive Oxygen Species (ROS) (*OCl, *HO⁻) utilizing electron and proton donation, ion and dissolved gas transport to produce a specific redox balanced set of molecules and ions. This redox-balanced set of molecules and ions in combination are a potent anti-infective and help the immune system identify and destroy malfunctioning cells.

This electrolyzed saline solution is then administered to a human or warm-blooded animal for therapeutic use. Preferably, the electrolyzed saline solution is administered by injection, oral or anal ingestion, applied topically, used as a bath, applied in a wound dressing, or inhaled in atomized form.

The container for producing the electrolyzed saline solutions is fabricated from a biologically compatible material. In addition, the anode is made of a base metal selected from the group consisting of platinum, niobium, titanium or any metal compatible with platinum bonding with an outer layer of platinum bonded to the base metal. The shape of the anode has a cylindrical, or flat (planar) shaped structure. The anode is preferably permeable to fluid flow.

Usually the cathode is positioned coaxially or in parallel in relation to the anode. This cathode is made of a base metal selected from the group consisting of platinum, niobium, titanium or any metal compatible with platinum bonding with an outer layer of platinum bonded to the base metal and has a cylindrical, or flat (planar) shaped structure similar to that of the anode and is also preferably permeable to fluid flow.

The spacing between the surfaces of the cathode and the anode is typically not greater than about one inch. This invention has means to circulate and regulate the temperature of fluids during production, has appropriate electrode design and has methods that effectively stabilize the composition of the resulting solution.

The temperature, fluid flow and effective voltage are chosen as to eliminate production of chlorates and to create the desired mixture of components. These parameters are determined by experimentation. The resulting solution is consistently stable and suitable for in vivo therapeutic applications. The stable ROS concentration, for example, has a variation of less than 5% from batch to batch and from device to device when the same set of parameters are employed by each.

The effective voltage may be applied by direct current, alternating current, or various combinations of alternating current and direct current power sources, resulting in a combined effective voltage ranging anywhere between 0 and 30 volts. The effective voltage is chosen to eliminate the production of chlorates and to create the desired mixture of components containing stable ROS. For example, a typical temperature range of the saline solution is from 30 deg. F. to 100 deg. F. In the lower temperature range, less O₂ is absorbed by the fluid and the fluid has smaller electrical conductivity, therefore higher effective voltages can be utilized to maintain adequate electrical current required to provide regulated amounts of stable ROS without significantly increasing the probability of creating chlorates and while maintaining a pH of 7.2 to 7.5.

The effective voltage may be adjusted, as desired, to regulate the concentration of the components and the pH of the resulting solution over a large variety of temperatures and fluid flows. Wherein it is difficult to theoretically determine the concentrations of all the various resulting chemical components when given any specific set of parameters, the optimal effective voltage, fluid temperature and flow are determined by experimentation. This methodology allows for the intentional regulation of concentrations of the specific chemical components in these stable ROS enriched solutions, allowing for the optimization of solutions intended for specific purposes.

The method and apparatus thus provides a stable, ROS enriched, antimicrobial, non-toxic electrolyzed saline solutions with a specific redox-balanced set of molecules and ions in solution, hereinafter referred to as “electrolyzed saline solution” (ESS) that has the ability to attack infective microbes and enhances the ability of the immune system to recognize and destroy damaged or malfunctioning cells. An example of a preferred embodiment of the ESS solution is that produced by Reoxcyn Discoveries Group, Inc. of Sandy, Utah under the tradename “Reoxcyn™”.

ESS solutions are balanced to normal and hypertonic saline and have been shown through extensive, repeatable research by accredited laboratories to be stable, nontoxic and exhibit remarkable antimicrobial, antiviral and therapeutic characteristics. Besides the therapeutic applications, the nature of these solutions also makes them suitable for applications in food safety, animal health, agriculture and sterilization. The ESS solutions exhibit a marked lack of toxicity upon intravenous, aspired, oral or topical application in mammals.

ESS solutions provide a broad platform for anti-infective and therapeutic applications covering several potential areas of use, including topical disinfection, antimicrobial application, wound treatment, oxidative stress reduction and enhancement of immune function. ESS solutions, being that they contain regulated amounts of stable reactive oxygen species (ROS), are particularly suited for enhancing the ability of the immune system to recognize and destroy damaged or malfunctioning cells. Such solutions can also be administered in a number of different ways appropriate for the desired therapeutic application.

Furthermore, all of the molecular components found in these solutions are involved in a growing field of scientific investigation categorized as redox messaging and regulation of genes. Such molecular components, being a balanced set of reduced species (RS) and reactive oxygen species (ROS), are the same molecules and ions that mirror those found in biological systems and are intimately involved in the ability of the immune system to recognize, detect, eliminate and heal infected, damaged or mutated tissues in mammals.

The measurement of concentrations of ROS inside the solutions has been done by means of a fluoro spectrometer, Nanodrop 3300, and three varieties of fluorescent dyes, R-Phycoerytherin (R-PE), Hydroxyphenyl fluorescein (HPF) and Aminophenyl fluorescein (APF), that are commonly used to determine relative ROS concentrations inside active biological systems and cells. The molecules in these dyes change shape, and therefore fluoresce only when exposed to molecular components in ROS. The resulting change in fluorescence can then be detected by the fluoro spectrometer and can be related to the concentration of ROS present. ROS concentrations in ESS solutions are verified and detected by either APF or R-PE fluorescent dyes, both of which produce entirely consistent measurements of relative concentrations of ROS in various concentrations and dilutions of ESS solutions. ROS measurements in ESS solutions have been linked using R-PE fluorescent dye, to the reaction of this dye to regulated concentrations of 2/2′-Axobis(2-methylpropionamide)dihidrochloride, a molecule that produces known amounts of ROS. This is not an absolute measurement, but it relates ROS in ESS to amounts of a known producer of ROS.

These fluorescent dyes are often used in combination with a fluorescence microscope to create high-resolution images of the build-up of ROS (oxidative stress) inside individual living cells. These dyes have been shown to specifically be sensitive to concentrations of ROS regardless of complex surrounding chemical environments.

Although APF and R-PE dyes are capable of measuring relative ROS concentrations in ESS solutions, no known absolute standard concentration for stabilized ROS in pure saline solutions exists. Furthermore, discrepancies in the decay time of these fluorescent dyes make measuring standardized amounts of ROS in other solutions incompatible with measuring those found in ESS. This may be due, in part, to the molecular complexes in ESS solutions that keep the ROS concentration stable, effectively shielding the free radicals from readily reacting with the dyes. The standard for ROS concentration in ESS solutions is therefore measured relative to the ROS concentration in a standardized solution that has been used in all of the antimicrobial and toxicity studies to date, both published and unpublished. Methods to measure absolute ROS concentrations in ESS solutions are actively being pursued.

The regulated amounts of ROS, thus measured, inside a variety of the ESS solutions produced by various embodiments of this invention have been shown to be stable, consistent and predictable, sufficient for therapeutic applications.

The method and apparatus thus produce a stable, non-toxic, antimicrobial electrolyzed saline solution with a broad range of anti-infective and therapeutic applications. The resulting electrolyzed saline solution is balanced to normal and hypertonic saline and has been shown to exhibit remarkable antimicrobial, antiviral and therapeutic characteristics. The nature of this electrolyzed saline solution makes it suitable for applications in food safety, animal health, agriculture and sterilization. These electrolyzed saline solutions also exhibit a marked lack of toxicity upon intravenous, aspired, oral or topical application in mammals. The therapeutic applications represent a broad platform, possibly covering a variety of potential areas of use, including topical disinfection, antimicrobial application, wound treatment, oxidative stress reduction and enhancement of immune function to better detect malfunctioning cells.

DESCRIPTION OF THE DRAWINGS

FIG. 1 is a side view of one preferred embodiment of the invention.

FIG. 2 is a top view of the preferred embodiment of the invention shown in FIG. 1.

DESCRIPTION OF THE ILLUSTRATED EMBODIMENTS

FIG. 1 is a side view of an embodiment of the invention. It has a container 101, which holds a saline solution having a saline concentration of at least about 0.15% to 1.0% by weight. The container may be fitted with a lid 100. The container 101 has a cylindrical anode 101 a and a surrounding concentric cylindrical cathode 101 b positioned on its bottom 105. The anode 101 a and cathode 101 b are operably associated with a power supply 107. The power supply 107 provides a source of electrical current with an effective voltage of under 30 volts via wires 106 affixed to the anode 101 a and a cathode 101 b.

The anode 101 a is a mesh cylindrical ring comprised of titanium with an outer layer of platinum bonded to the titanium base. The cathode 101 b is a cylindrical mesh ring comprised of titanium with an outer layer of platinum bonded to the titanium base that is positioned coaxially about the anode 101 a. The spacing between the cathode 101 b and the anode 101 a, at the preferred flow rate below, is typically not greater than about one inch. Moreover, the effective voltage potential between the cathode 101 b and the anode 101 a is not greater than a preferred amount, typically under 30 volts.

A temperature regulation device, such as a combination heating/cooling device, is positioned along the sides 104 inside the container 101 to exchange heat with the saline solution in order to maintain the saline solution at a desired temperature between 30 deg. F. to 100 deg. F.

A circulation tube 102 is mounted on the exterior of the container 101 with openings connecting and in communication with the top and bottom interior of the container 101. The circulation tube 102 is associated with a fluid pump 103 to provide for fluid circulation and flow inside the container 101. This allows saline solution in the container 101 to flow through the anode 101 a and cathode 101 b assembly at a preferred flow rate, typically between 0.1 to 50 cc/cm²/sec.

FIG. 1 also shows a second circulation tube 102 and fluid pump 103 similarly structured and mounted on the exterior of the opposite side of the container 101 that performs a similar fluid circulation function. This two tube 102 circulation structure and flow pattern insures complete mixing and electrolysis of the saline solution to produce ROS concentrations calculated to be between 0.05 and 50 ppm.

FIG. 2 is a top view of the preferred embodiment of the invention shown in FIG. 1.

Although this reference has made reference to the illustrated embodiments, it is not intended to limit the scope of the claims. The claims themselves recite those features deemed essential to the invention. 

We claim:
 1. A stable, non-toxic, antimicrobial electrolyzed saline solution exhibiting anti-infective and immune-enhancing potential as a therapeutic containing regulated amounts of stable reactive oxygen species (ROS), comprising: a first target mixture of chemically reduced and oxidized species including Hypochlorous acid (HOCl), Hypochlorites (OCl⁻, NaClO), dissolved Oxygen (O₂), Chlorine (Cl₂) between 1 to 200 ppm and Hydrogen (H₂) gas, Hydrogen Peroxide (H₂O₂), Hydrogen ions (H⁺), Hypochloride (ClO) and corresponding amounts of Superoxides (*O₂ ⁻, HO₂), Ozone (O₃) from 1 to 50 ppm, Activated Hydrogen ions (H), Chloride ions (Cl⁻), Hydroxides (NaOH, OH), Singlet Oxygen (*O₂) and other forms of Reactive Oxygen Species (ROS) (*OCl, *HO⁻), and wherein total ROS is between about 0.05 and about 50 ppm, wherein the first target mixture is configured to mirror a known biological system including a second target mixture of chemically reduced and oxidized molecules.
 2. The electrolyzed saline solution of claim 1, wherein the first target mixture and the second target mixture are measured using a fluorospectrometer configured to measure concentrations of reactive oxygen species.
 3. The electrolyzed saline solution of claim 2, wherein the concentrations are measured using at least one fluorescent dye selected from R-phycoerytherin, hydroxyphenyl fluorescein, and aminophenyl fluorescein.
 4. The electrolyzed saline solution of claim 1, wherein the first target mixture of chemically reduced and oxidized molecules includes at least *O₂.
 5. The electrolyzed saline solution of claim 1, wherein the electrolyzed saline solution is stored in a container is fabricated from a biologically compatible material.
 6. An electrolyzed saline solution comprising: a first target mixture of chemically reduced and oxidized species, wherein the first target mixture is configured to mirror a known biological system including a second target mixture of chemically reduced and oxidized molecules.
 7. The electrolyzed saline solution of claim 6, wherein the first target mixture and the second target mixture are measured using a fluorospectrometer configured to measure concentrations of reactive oxygen species.
 8. The electrolyzed saline solution of claim 7, wherein the concentrations are measured using at least one fluorescent dye selected from R-phycoerytherin, hydroxyphenyl fluorescein, and aminophenyl fluorescein.
 9. The electrolyzed saline solution of claim 6, wherein the first target mixture of chemically reduced and oxidized molecules includes at least one of HOCl, OCl⁻, NaClO, O₂, H₂, H₂O₂, H⁺, ClO, *O₂ ⁻, HO₂, O₃, H⁻, Cl⁻, NaOH, OH⁻, *O₂, *OCl, or *HO⁻.
 10. The electrolyzed saline solution of claim 6, wherein Cl₂ is present between about 1 ppm and about 200 ppm.
 11. The electrolyzed saline solution of claim 6, wherein O₃ is present between about 1 ppm and about 50 ppm.
 12. The electrolyzed saline solution of claim 6, wherein total stable ROS compounds are present between about 0.05 ppm and about 50 ppm.
 13. The electrolyzed saline solution of claim 6, wherein the electrolyzed saline solution is stored in a container is fabricated from a biologically compatible material.
 14. An electrolyzed saline solution comprising: a first target mixture of chemically reduced and oxidized species, wherein the first target mixture is configured to mirror a known biological system including a second target mixture of chemically reduced and oxidized molecules by measuring concentrations of reactive oxygen species using a fluorospectrometer and at least one fluorescent dye selected from Rphycoerytherin, hydroxyphenyl fluorescein, and aminophenyl fluorescein.
 15. The electrolyzed saline solution of claim 14, wherein the first target mixture of chemically reduced and oxidized molecules includes at least one of HOCl, OCl⁻, NaClO, O₂, H₂, H₂O₂, H⁺, ClO, *O₂ ⁻, HO₂, O₃, H⁻, Cl⁻, NaOH, OH⁻, *O₂, *OCl, or *HO⁻.
 16. The electrolyzed saline solution of claim 15, wherein Cl₂ is present between about 1 ppm and about 200 ppm.
 17. The electrolyzed saline solution of claim 15, wherein O₃ is present between about 1 ppm and about 50 ppm.
 18. The electrolyzed saline solution of claim 15, wherein total stable ROS compounds are present between about 0.05 ppm and about 50 ppm.
 19. The electrolyzed saline solution of claim 14, wherein the electrolyzed saline solution is stored in a container is fabricated from a biologically compatible material. 